Scanning Electrochemical Microscopy of DNA Monolayers Modified with Nile Blue

Scanning electrochemical microscopy (SECM) is used to probe long-range charge transport (CT) through DNA monolayers containing the redox-active Nile Blue (NB) intercalator covalently affixed at a specific location in the DNA film. At substrate potentials negative of the formal potential of covalently attached NB, the electrocatalytic reduction of Fe(CN)63− generated at the SECM tip is observed only when NB is located at the DNA/solution interface; for DNA films containing NB in close proximity to the DNA/electrode interface, the electrocatalytic effect is absent. This behavior is consistent with both rapid DNA-mediated CT between the NB intercalator and the gold electrode as well as a rate-limiting electron transfer between NB and the solution phase Fe(CN)63−. The DNA-mediated nature of the catalytic cycle is confirmed through sequence-specific and localized detection of attomoles of TATA-binding protein, a transcription factor that severely distorts DNA upon binding. Importantly, the strategy outlined here is general and allows for the local investigation of the surface characteristics of DNA monolayers both in the absence and in the presence of DNA binding proteins. These experiments highlight the utility of DNA-modified electrodes as versatile platforms for SECM detection schemes that take advantage of CT mediated by the DNA base pair stack

Revealing ensemble state transition patterns in multi-electrode neuronal recordings using hidden Markov models

In order to harness the computational capacity of dissociated cultured neuronal networks, it is necessary to understand neuronal dynamics and connectivity on a mesoscopic scale. To this end, this paper uncovers dynamic spatiotemporal patterns emerging from electrically stimulated neuronal cultures using hidden Markov models (HMMs) to characterize multi-channel spike trains as a progression of patterns of underlying states of neuronal activity. However, experimentation aimed at optimal choice of parameters for such models is essential and results are reported in detail. Results derived from ensemble neuronal data revealed highly repeatable patterns of state transitions in the order of milliseconds in response to probing stimuli

Verapamil protects against progression of experimental chronic renal failure

Verapamil protects against progression of experimental chronic renal failure. Chronic administration of verapamil (Ver) decreases nephrocalcinosis and tubular ultrastructural abnormalities in the remnant model of chronic renal disease. In the present study, the effect of chronic Ver administration on renal function, renal histology and mortality after subtotal nephrectomy was examined. Fourteen days after staged subtotal nephrectomy rats were paired according to renal functional impairment, mean arterial pressure (MAP), and body weight. Rats were pair fed and received either Ver (0.1 µg/g sc bid, N = 10) or saline (0.1ml sc bid, N = 10) for up to 23 weeks. Both members of each pair were sacrificed shortly before the uremic death of controls. At sacrifice, rats treated with Ver had a lower serum creatinine (2.29 vs. 2.99 mg/dl, P < 0.05) and a higher creatinine clearance (318 vs. 164 µl/min, P < 0.05) than controls. In a second experiment, survival was superior in rats treated with Ver than in controls from week seven (P < 0.0025 by week 14). Serum creatinine was higher at week 10 in control rats (1.68 vs. 1.10 mg/dl, P < 0.05). MAP was no different between the two groups, irrespective of the time between Ver administration and the measurement of MAP. Histological damage and nephrocalcinosis were worse, and renal and myocardial calcium content was higher in controls. In conclusion, independent of any effect on systematic MAP, chronic administration of Ver protects against renal dysfunction, histological damage, nephrocalcinosis and myocardictl calcification, and improves survival in the remnant model of chronic renal disease

Heart rate variability during high-speed treadmill exercise and recovery in Thoroughbred racehorses presented for poor performance

Background: Heart rate variability (HRV) analysis measures the inter-beat interval variation of successive cardiac cycles. Measurement of these indices has been used to assess cardiac autonomic modulation and for arrhythmia identification in exercising horses.Objectives: To report HRV indices during submaximal exercise, strenuous exercise and recovery, and explore relationships with clinical conditions (arrhythmias, lameness, equine gastric ulcer syndrome (EGUS), lower airway inflammation and upper respiratory tract obstructions (URTO) in Thoroughbred (TB) racehorses.Study Design: Retrospective, observational cross-sectional study.Methods: One hundred and eighty Thoroughbred horses underwent a treadmill exercise test with simultaneous electrocardiographic (ECG) recording. Time-domain HRV indices (standard deviation of the R-R interval (SDRR); root mean square of successive differences (RMSSD)) were derived for submaximal and strenuous exercise and recovery segments. Clinical conditions (arrhythmia (during each phase of exercise), lameness, EGUS, lower airway inflammation and URTO) were assigned to binary categories for statistical analysis. Relationships between selected HRV indices and the clinical conditions were explored using linear regression models. Results: During submaximal exercise, lameness was associated with decreased logRMSSD (B= -0.19 95% CI -0.31 to -0.06, P= 0.006) and arrhythmia was associated with increased logRMSSD (B= 0.31 95% CI 0.01-.608, P=0.04). During strenuous exercise, arrhythmia was associated with increased HRV indices (logSDRR B= 0.51 95% CI 0.40-0.62, PMain Limitations: The main limitations of this retrospective study were that not every horse had the full range of clinical testing, therefore some horses may have had undetected abnormalities. Conclusions: The presence of arrhythmia increased HRV in both phases of exercise and recovery. Lameness decreased HRV during submaximal exercise

Shoot yield drives phosphorus use efficiency in Brassica oleracea and correlates with root architecture traits

The environmental and financial costs of using inorganic phosphate fertilizers to maintain crop yield and quality are high. Breeding crops that acquire and use phosphorus (P) more efficiently could reduce these costs. The variation in shoot P concentration (shoot-P) and various measures of P use efficiency (PUE) were quantified among 355 Brassica oleracea L. accessions, 74 current commercial cultivars, and 90 doubled haploid (DH) mapping lines from a reference genetic mapping population. Accessions were grown at two or more external P concentrations in glasshouse experiments; commercial and DH accessions were also grown in replicated field experiments. Within the substantial species-wide diversity observed for shoot-P and various measures of PUE in B. oleracea, current commercial cultivars have greater PUE than would be expected by chance. This may be a consequence of breeding for increased yield, which is a significant component of most measures of PUE, or early establishment. Root development and architecture correlate with PUE; in particular, lateral root number, length, and growth rate. Significant quantitative trait loci associated with shoot-P and PUE occur on chromosomes C3 and C7. These data provide information to initiate breeding programmes to improve PUE in B. oleracea

Biological methods to assess unaccompanied asylum-seeking children's age

Report by the interim Age Estimation Science Advisory Committee (AESAC) on scientific methodologies for assessing the age of unaccompanied asylum-seeking children

Biological methods to assess unaccompanied asylum-seeking children's age

Report by the interim Age Estimation Science Advisory Committee (AESAC) on scientific methodologies for assessing the age of unaccompanied asylum-seeking children

Biological methods to assess unaccompanied asylum-seeking children’s age:Interim Age Estimation Science Advisory Committee

Report by the interim Age Estimation Science Advisory Committee (AESAC) on scientific methodologies for assessing the age of unaccompanied asylum-seeking children.<br/

Biological methods to assess unaccompanied asylum-seeking children’s age:Interim Age Estimation Science Advisory Committee

Report by the interim Age Estimation Science Advisory Committee (AESAC) on scientific methodologies for assessing the age of unaccompanied asylum-seeking children.<br/

Acute phosphate depletion and in vitro rat proximal tubule injury: Protection by glycine and acidosis

Acute phosphate depletion and in vitro rat proximal tubule injury: Protection by glycine and acidosis.The effects of phosphate (PO4) removal from Krebs Henseleit buffer on freshly isolated rat proximal tubules (rPT) were assessed by measuring Ca2+ uptake (nmol/mg protein), cellular adenosine triphosphate (ATP) (nmol/mg), tissue K+ content (nmol/mg) and lactate dehydrogenase (LDH) as an index of cell integrity. Ca2+ uptake increased by 50% in rPT incubated in zero PO4 medium as compared to control (2.6 ±0.1 vs. 3.9 ±0.19, P < 0.001) and LDH release increased 2.5-fold from 14.2 ±0.6 to 31.6 ±1.6%, P < 0.001. Neither verapamil (200 µM) nor mepacrine (50 µM) reduced Ca2+ uptake or decreased LDH release suggesting that the increased Ca2+ uptake was not occurring through potential operated channels and that phospholipase-induced cell injury was not the cause of increased LDH release. Either glycine (2 mM) or extracellular fluid acidosis (pH 7.06), however, significantly diminished rPT injury and Ca2+ uptake. Specifically, as compared to the increased LDH released in untreated, PO4-depleted rPT, LDH release was diminished significantly by glycine treatment (31.0 ±0.9 vs. 15.5 ±1.6%, P < 0.001) or acidosis (30.3 ±0.04 vs. 19.2 ±0.9%, P < 0.01). Ca2+ uptake did not increase in glycine treated tubules (2.6 ±0.1 vs. 2.8 ±0.2 nmol/mg, NS) or in the presence of acidosis (2.6 ±0.1 vs. 2.97 ±0.17 nmol/mg, NS). ATP concentrations were markedly reduced by PO4 depletion (2.8±0.2 vs. 4.8±0.3 nmol/mg, P < 0.001) and remained at low levels during either acidosis or glycine-induced protection. ATP depletion was accompanied by loss of K+ from rPT and this was only modestly attenuated by either glycine or acidosis. Total cell PO4 was not significantly altered, however, perchloric acid (PCA) extractable free PO4 was reduced significantly (33.3 ±4.5 to 15.9 ±3.5 nmol/mg, P < 0.01). The rPT injury, associated with acute PO4 depletion, may be related to Ca2+ uptake since Ca2+ uptake and LDH release were both attenuated by glycine administration or acidosis